J. Braz. Chem. Soc., Vol. 00, No. 00, 1-4, 2010.

Printed in Brazil - ©2010 Sociedade Brasileira de Química

0103 - 5053 $6.00+0.00

Short Report
New Isoflavone Derivative and others Flavonoids from the Resin of
Amburana cearensis

Paulo N. Bandeira,*, a Silvana S. de Farias,a Telma L. G. Lemos,b Raimundo Braz-Filho,c

Hélcio S. Santos,a Maria R. J. R. Albuquerquea and Sônia M. O. Costad
a
Laboratório de Química Orgânica, Universidade Estadual Vale do Acaraú,
62040-370 Sobral-CE, Brazil
b
Departamento de Química Orgânica e Inorgânica, Universidade Federal do Ceará,
60451-970 Fortaleza-CE, Brazil
c
Laboratório de Ciências Químicas, CCT, Universidade Estadual do Norte Fluminense Darcy
Ribeiro, 28013-602 Campos-RJ, Brazil
Departamento de Física e Química, Universidade Estadual do Ceará,
d

60749-000 Fortaleza-CE, Brazil

A investigação fitoquímica da resina de Amburana cearensis A. C. Smith permitiu o

isolamento de um novo composto: 3’,4’-dimetóxi-1’-(7-metóxi-4-oxo-4H-cromen-3-il)
benzo-2’,5’-quinona (1), juntamente com seis compostos conhecidos e identificados
como: 4,2’,4’-triidroxichalcona (2), 7,8,3’,4’-tetrametoxiisoflavona (3), 4,2’,4’-triidroxi-3-
metoxichalcona  (4), 3,4,5-trimetoxicinamaldeido (5), 3’,4’-dimetoxi-7-hidroxiisoflavona (6) e
6,7,4’-trimetoxi-3’-hidroxiisoflavona (7). As estruturas foram estabelecidas com base na análise
dos dados espectrométricos de: IV, EI-EM, HR-ESI-EM e RMN incluindo experimentos 2D.

Phytochemical investigation of the resin of Amburana cearensis A. C. Smith

allowed the isolation of a new compound: 3’,4’-dimethoxy-1’-(7-methoxy-4-oxo-4H-
cromen-3-yl)benzo-2’,5’-quinone (1), together with six known compounds identified as:
4,2’,4’-trihydroxychalcone  (2), 7,8,3’,4’-tetramethoxyisoflavone (3), 4,2’,4’-trihydroxy-3-
methoxychalcone (4), 3,4,5-trimethoxycinnamaldehyde (5), 3’,4’-dimethoxy-7-hydroxyisoflavone
(6) and 6,7,4’-trimethoxy-3’-hydroxyisoflavone (7). The structures were established from the IR,
HR‑ESI‑MS and NMR spectral data, including 2D-NMR experiments.

Keywords: Amburana cearensis, resin, phytochemical investigation, flavonoids

Introduction kaempferol and polyphenols isolated from the trunk bark

Amburana cearensis A. C. Smith (Sin. Torrensea
cearensis Fr. AlI), Leguminoseae family, Papilionoideae
(Fabaceae), is a tree popularly known as “imburana de
cheiro”, “cerejera” and “cumaru” in northeast of Brazi1.1,2
The syrup of trunk bark is used in folk medicine for
treatment of respiratory diseases as cold, bronchits and
asthma. Pharmacological studies using hydroalcoolic syrup
from the bark trunk showed anti-inflammatory, analgesic,
bronchodilator and antinociceptive activities.3-6 Recent
studies7 have shown that amburoside A, isokaempferide,
*e-mail: bandeirapn@yahoo.com.br
of A. cearensis also showed present neuroprotective,
antioxidant, antiinflamatory, anticarcinogenic and
hepatoprotective activities.
Previous studies with the bark from A. cearensis
resulted in the isolation of 4-(O-b-D-glucopyranosyl)-
hydroxy-7-(3’,4’-dihydroxy-benzoyl)-benzyl, 4-(O-b-D-
glucopyranosyl)-hydroxy-7-(3’-methoxy-4’-hydroxy-
benzoyl)-benzyl, coumarin, sucrose, vanilic acid,
protocatechuic acids, afrormosin, isokaempferide,
kaempferol, quercetin, 4’ -methoxy-fisetin, amburoside A
and a mixture of b-sitosterol and stigmasterol glucoside).8,9
Phytochemical investigation was carried out recently with
especies obtained from seed germinations.10
2 New Isoflavone Derivative and others Flavonoids from the Resin of Amburana cearensis J. Braz. Chem. Soc.

This paper describes the chemical investigation of the voucher (No. 847) specimen is deposited in the Herbarium
resin of A. cearensis, which resulted in the isolation of a Prisco Bezzera of this University.
new compound named as 3’,4’-dimethoxy-l’-(7-methoxy-
4-oxo-4H-cromen-3-yl)benzo-2’,5’-quinone (1). Extraction and isolation

Experimental The resin (400 g) was submitted to an extraction

General procedure
The melting point was determined using a Mettler
Toledo FP82HT micromelting point apparatus. The IR
spectra were measured in KBr pellets, using Perkin-Elmer,
FT-IR Spectrum 1000. All the NMR data were recorded
using a Bruker Avance DPX 300 and Avance DRX-500
spectrometer operating in the frequency of the hydrogen
at 300.13 and 500.13 MHz in the frequency of the carbon
for 75.47 and 125.75 MHz. The spectra were recorded
in CDCl3, C5D5N and CD3OD. High resolution data were
obtained in a MS-IT-TOF mass spectrometer.
with EtOH at room temperature and yielded (23.46 g).
The EtOH extract was dissolved in 200 mL of water
and partitioned in the CHCl3 and EtOAc yielded (8.1 g)
and (0.7 g) respectively. The EtOAc fraction (0.7 g)
was subjected to CC on SiO2 using EtOAc, CHCl3 and
MeOH. The CHCl3 fraction (0.15 g) was chromatographed
in sephadex LH-20 column using MeOH, obtaining
compound 1 (18 mg) and 2 (30 mg). The CHCl3 fraction
(8.1 g) was subjected to CC on SiO2 using EtOAc, CHCl3
and MeOH. The CHCl3 fraction (0.30 g) was successively
chromatographed in sephadex LH-20 column using
MeOH and obtaining compound 3 (40 mg), 4 (23 mg),
5 (15 mg), 6 (10 mg) and 7 (16 mg).

Plant material 3’,4’-Dimethoxy-1’-(7-methoxy-4-oxo-4H-cromen-3-yl)

The resin of A. cearensis was collected in city of
Limoeiro do Norte, Ceará, Brazil. The plant material was
identified in the Departamento de Biologia do Centro de
Ciências da Universidade Federal do Ceará, Brazil. A
benzo-2’,5’-quinone (1)
Orange solid, mp 196-198 oC; IR (KBr) νmax/cm-1: 1649,
1628, 1595, 1439, 1279, 1100, 835; 1H and 13C NMR (see
Table 1); HR-ESI-MS spectrum revealed formation of
molecular ions m/z 343.0931 (C18H14O7 + H+), 365.0661

Table 1. 1H NMR (500 MHz) and 13C NMR (125 MHz) data of 1 (in CDCl3) compared with values of 8 (DMSO-d6, (dd, ppm), J in Hz*

1 8
1
H,13C-HMQC 1
H,13C-HMBC
d (C) d (H) 2
J 3
J d (C) d (H)
C(2) 156.5 8.16 (s) 157,5 8.36 (s)
C(3) 116.3 H-2 H-6’ 115.5
C(4) 174.4 H-2; H-5 178.8
C(5) 127.9 8.15 (d, J 8.9) 161.6
C(6) 115.1 7.02 (dd, J 8.9, 2.3) H-8 99.5 6.44 (d, J 1.5)
C(7) 165.0 H-6; H-8 MeO-7 164.7
C(8) 100.4 6.88 (d, J (2.3) H-6 94.1 6.27 (d, J 1.5)
C(9) 158.0 H-8 H-2 157.1
C(10) 118.0 H-6; H-8 104.0
C(1’) 135.0 H-6’ H-2 137.9
C(2’) 183.9 H-6’; MeO-2’ 185.0
C(3’) 145.0 MeO-3’ 107.9 6.26 (s)
C(4’) 158.2 MeO-4’; H-6’ 158.4
C(5’) 183.9 181.4
C(6’) 133.9 7.17(s) 133.2 7.05 (s)
MeO-7 55.9 3.93 (s)
MeO-3’ 61.2 4.03 (s)
MeO-4’ 61.5 4.08 (s) 56.3 3.83 (s)
*Number of hydrogens bound to carbon atoms deduced by comparative analysis of 1H and DEPT 13C NMR spectra. Chemical shifts and coupling constants
(J) were obtained of 1D 1H NMR spectrum. 1H,1H COSY and 1H, 1H NOESY experiments were also used for these assignments in compared with literature
data 8 (ref. 13).
Vol. 00, No. 00, 2010
(C18H14O7 + Na+) and 381.0438 (C18H14O7 + K+ ). LR-EI-
MS m/z (rel; Int.): ([M+], 94.5%), 200 [M+ - 142] 100%,
150 [M+ - 193] 32.9%, 327 [M+ - 15] 8.2%.
Results and Discussion
Compound 1 was obtained as orange solid. The HR-ESI-
MS spectrum showed a pseudo molecular íons peaks at m/z
343.0931 (C18H14O7 + H+, requires 343.0818), 365.0661
(C18H14O7 + Na+ , requires 365.0637) and 38l.0438 (C18H14O7
+ K+ , requires 38l.0377). The IR spectrum exhibited bands
at 1650, 1628 and 1595 (nc=0) cm-1 suggesting the presence of
three conjugated carbonyl groups.11,12 Comparative analysis
of the 1H and DEPT l3C NMR spectra (Table 1) allowed to
recognize signals for three methoxyl groups and fifteen
sp2 carbons: five methines (including one linked to oxygen
atom at dc 156.5, compatible with CH-2 of isoflavones) and
ten non-hydrogenated [including three carbonyl groups at
dc 174.4 (C-4), dc 183.9 (C-2’ and C-5’) and four linked
to oxygen atoms: dc 165.0 (C-7), dc 158.0 (C-9), dc 145.0
Figure 1. Structure of the chemical constituents (1-7) isolated of resin of Amburana cearensis A.C Smith and the compound 8 from reference 13.
Bandeira et al. 3
(C-3’) and dc 158.2 (C-4’)]. The signals observed in the
13
C NMR spectra at dc 183.9 and 174.4 were attributed to
carbonyl groups at the 1,4-benzoquinone and flavonoid
moieties,13 respectively. Furthermore, the 1H NMR exhibited
a characteristic a singlet at dH 8.16, assigned to H-2 of an
isoflavone, confirmed by HMQC spectrum by the presence
of a transversal peak corresponding to direct correlation of
this hydrogen signal with l3C signal of methine at dc 156.5
(CH‑2), and at dH 3.93, dH 4.03 and dH 4.08, corresponding
to three methoxyl groups located at C-7, C-3’ and C-4’,
respectively. The location of these methoxyl groups was
observed at the heteronuclear long range correlations in the
HMBC spectrum (Table 1), which showed correlations of
the methoxyl signal at dH 3.93 with C-7 (dc 165.0, 3JCH, dH
4.03 (MeO-3’) with dc 145.0 (C-3’, 3JCH) and dH 4.08 (MeO-
4’) with dc 158.2 (C-4’, 3JCH). The presence of a methoxyl
group at C-7 was additionally confirmed by nOe effects
observed in the 1H, 1H NOESY spectrum, which revealed
spatial interactions with the hydrogen atoms H-6 and H-8.
The multiplicities of these signals of hydrogen atoms H-6
4 New Isoflavone Derivative and others Flavonoids from the Resin of Amburana cearensis
(dH 7.02, dd, J 8.9 and 2.3 Hz) and H-8 (dH 6.88, d, J 2.3 Hz)
in combination with the corresponding H-5 (dH 8.15, d,
J 8.9 Hz), correlated via one bond in the HMQC spectrum
with the 13C signal at 127.9, allowed to define the ring A,
sustaining a methoxyl group at carbon atom C-7. The HMBC
spectrum was also used to characterize the presence of a
2,3-dimethoxy-1,4- benzoquinone moiety at C-3 through
heteronuclear long range correlation (Table 1) of the carbon
C-4’ (dc 158.2), with both H-6’ (dH 7.17, s, 3JCH) and MeO‑4’
(dH 4.08, s, 3JCH), C-3’ (dc 145.0) with MeO-3’ (dH 4.03, s,
3
JCH), C-1’ (dH 135.0) with both H-2 (dH 8.16, s, 3JCH) and
H-6’ (dH 7.17, s, 2JCH) and C-3 (dc 116.3) with both H-2 (dH
8.16, s, 2JCH) and H-6’ (dH 7.17, s, 3JCH). Additional analysis
of the l3C NMR spectra of 1 revealed, the l3C chemical
shifts (dc 145.0) for the C-3’ and (dc 158.2) for the C-4’
and for the C-2’/C-5’ (dc 183.9), whereby after comparison
of these said data with the correspondent values, dc 107.9
(CH-3’), dc 158.4 (C-4’), dc 185.0 (C-2’) and dc 181.4 (C‑5’),
described in the literature,13 suggests the presence of an
additional methoxyl group at carbon C-3’ of 1 as differences
observed between spectral data of 1 and of 8 (Table 1). The
base peak at m/z 200 (100%) observed in the LR-EI-MS,
significantly contributed to characterize and to locate the
2,3-dimethoxy-l,4- benzoquinone moiety at C-3 (Figure 1).
Therefore, the structure of compound 1 was assigned
as a new isoflavone and it is characterized as 3’,4’-
dimethoxy-1’-(7 -methoxy-4-oxo-4H-cromen-3-yl)-benzo-
2’,5’-quinone.
The compounds 2, 3, 4, 5, 6 and 7 were identified
through spectroscopic data compared with literature data
as: 4,2’,4’-trihydroxychalcone,14 7,8,3’,4’-tetrarnethoxy­
isoflavone,15 4,2’,4’-trihydroxy-3-methoxychalcone,16
3,4,5- trimethoxycinnamaldehyde,17 3’,4’-dimethoxy-7-
hydroxyisoflavone18 and 6,7,4’-trimethoxy- 3’-hydroxy­
isoflavone19 (Figure 1) , respectively.
Supplementary Information
Supplementary data are available free of charge at
http://jbcs.sbq.org.br, as PDF file.
Acknowledgments
The authors whishes to thank CNPq/FUNCAP for
financial support, CENAUREMN by NMR spectra and
LEMANOR by mass spectrum.
References
1. Corrêa, M. P.; Dicionário das Plantas Úteis do Brasil e das
Exóticas Cultivadas, Ministério da Agricultura: Brasília, 1984.
J. Braz. Chem. Soc.
2. Maia, G. N.; Caatinga: Árvores e Arbustos e Suas utilidades,
D & Z ed.: São Paulo, 2004.
3. Leal, L. K. A. M.; Matos, M. E.; Matos, F. J. A.; Ribeiro, R. A.;
Ferreira, F. V.; Viana, G. S. B.; Phytomedicine 1997, 4, 221.
4. Leal, L. K. A. M.; Nechio, M.; Silveira, E. R.; Canuto, K. M.;
Fontenele, J. B.; Ribeiro, R. A.; Viana, G. S. B.; Phytother. Res.
2003, 17, 335.
5. Oliveira, R. R. B.; Góis, R. M. O.; Siqueira, R. S.; Almeida, J.
R. G. S.; Lima, J. T. L.; Nunes, X. P.; Oliveira, V. R.; Siqueira,
J. S.; Quintons-Junior, L. J.; Braz. J. Pharmcog. 2009, 19, 672.
6. Leal, L. K. A. M.; Ferreira, A. A. G.; Bezerra, G. A.; Matos, F.
J. A.; Viana, G. S. B.; J. Ethnopharmacol. 2000, 70, 151.
7. Leal, L. K. A. M.; Canuto, K. M.; Costa, K. C. S.; Nobre-
Junior, H. V.; Vasconcelos, S. M.; Silveira, E. R.; Ferreira, M.
V. P.; Fontenele, J. B.; Andrade, G. M.; Viana, G. S. B.; Basic
Clin. Pharmacol. Toxicol. 2008, 104, 198; Leal, L. K. A. M.;
Nobre-Junior, H. V.; Cunha, G. M. A.; Moraes, M. O.; Pessoa,
C.; Oliveira, R. A.; Silveira, E. R.; Canuto, K. M.; Viana, G. S.
B.; Neurosci. Lett. 2005, 388, 86; Leal, L. K. A. M.; Costa, M.
F.; Pitombeira, M.; Barroso, V. M.; Silveira, E. R.; Canuto, K.
M.; Viana, G. S. B.; Life Sci. 2006, 79, 98; Costa-Lotufo, L.
V.; Jimenez, P. C.; Wilke, D. V.; Leal, L. K. A. M.; Cunha, G.
M. A.; Silveira, E. R.; Canuto, K. M.; Viana, G. S. B.; Moraes,
M. E. A,; de Moraes, M. O.; Pessoa, C.; J. Biosci. 2003, 58, 9;
Leal, L. K. A. M.; Fonseca, F. N.; Pereira, F. A.; Canuto, K. M.;
Felipe, C. F. B.; Fontenele, J. B.; Pitombeira, M. V.; Silveira,
E. R.; Viana, G. S. B.; Planta Med. 2008, 74, 497.
8. Bravo, J. A. B.; Sauvain, M.; Gimenez, A.; Muñoz, V.; Callapa,
J.; Le Men-Oliver, L.; Massiot, G.; Lavaud, C.; Phytochemistry
1999, 50, 71.
9. Canuto, K. M.; Silveira, E. R.; Quim. Nova 2006, 29, 1241.
10. Canuto, K. M.; Silveira, E. R.; Bezerra, A. M. E.; Quim. Nova
2010, 33, 662.
11. Silverstein, R. M.; Webster, F. X.; Identificação Espectrométrica
de Compostos Orgânicos, 6th ed., LTC, ed.: Rio de Janeiro, 2000.
12. Marques, W. B.; Santos, H. S.; Pessoa, O. D. L.; Braz-Filho,
R.; Lemos, T. L.G.; Phytochemistry 2000, 55, 793.
13. Hamburger, M. O.; Cordell, G. A.; J. Nat. Prod. 1987, 50, 696.
14. Markham, K. R.; Ternai, B.; Tetrahedron 1976, 32, 2607.
15. Rao, V.; Murthy, S. R.; Ward, R. T.; Phytochemistry 1984, 23,
1493.
16. Nielsen, S. F.; Christensen, S. B.; Cruciani, G.; Kharazmi, A.;
Liljefors, T.; J. Med. Chem. 1998, 41, 4819.
17. Mohammad, I.; Watterman, P. G.; J. Nat. Prod. 1985, 48, 328.
18. Rong, H.; Stevens, J. F.; Deizer, M. L.; De Cooman, L.; De
Keukeleire, D.; Planta Med. 1998, 64, 620.
19. Razdan, T. K.; Kachroo, P. K.; Qadri, A. K.; Kalla, A. K.;
Taneja, S. K.; Dhar, K. L.; Phytochemistry 1996, 41, 947.
Submitted: November 21, 2009
Published online: October 7, 2010